TUBERCULOUS SPUTUM 335 



incubator for one hour. To 10 c.c. of the homogeneous mixture add 1.5 c.c. of a 

 solution made up of one part chloroform and nine parts alcohol. Shake violently 

 and centrifuge for 15 minutes. Mix the sediment wit)/ egg albumin, smear out and 

 stain. 



When it is desired to culture the tubercle bacilli mix 20 c.c. of sputum with 65 c.c. 

 sterile water and add 15 c.c. antiformin. Stir the mixture with a glass rod. After 

 30 minutes to two hours we should have a homogeneous mixture. Centrifuge for 

 15 minutes or longer, wash the sediment twice with sterile salt solution and smear out 

 the well-washed sediment over serum or glycerine egg slants. The tubes should be 

 covered with black paper and the plugs paraffined. It must be remembered that for 

 culturing tubercle bacilli we must protect the growth from sunlight as this will kill 

 the organism. If fluid culture media are inoculated the transferred material should 

 be deposited on the surface. Should the particle sink growth will not occur. 



Sputum smears stained by some Romanowsky method or by the haematoxylin- 

 eosin stain are best adapted for the study of various cells, and in particular of the 

 eosinophile cells so characteristic of bronchial asthma. In sputum from cancer of 

 the lungs the large vacuolated cells may be found. 



When examining the sputum of the bronchopneumonia of influenza the formol 

 fuchsin gives the best results. The influenza bacilli are found in little masses, fre- 

 quently grouped about small collections of M. tetragenus. The cocci stain a rich 

 purplish-red, while the small influenza bacilli take on a light pink color. 



T. B. sputum showing a mixed infection with streptococci or pneumococci or with 

 the influenza bacillus makes for a bad prognosis. M. tetragenus, which often is 

 present when cavities exist, does not seem to be so unfavorable prognostically. 



Red cells show up well in specimens stained by the Romanowsky method; if 

 rouleaux formation is marked, it may indicate pulmonary infarction. 



In culturing sputum a mucopurulent mass should be washed in 

 sterile water and should then be dropped into a tube of sterile bouillon. 

 With a sterile swab it should be emulsified and successive streaks made 

 along the surface of an agar or glycerine agar plate. In obtaining cul- 

 tures from influenza sputum, first smear the material thoroughly over 

 a blood-serum slant; then inoculate, by thorough smearing over the 

 surface of successive blood-stieaked agar slants, the material on the 

 surface of the blood-serum slant. The platinum loop should be trans- 

 ferred from one slant to another without recharging. The influenza 

 bacillus seems to grow better if the blood-streaked agar slants are 

 prepared just before inoculating with the sputum. All that is necessary 

 is to sterilize an ear, puncture and take up the exuding blood with a 

 large loop. Cultures for tubercule bacilli are impracticable except with 

 antiformin. A guinea-pig should be inoculated. 



The blood-stained watery sputum of plague pneumonia should be cultured on 



